Protein arginine methyltransferase 6 is a novel substrate of protein arginine methyltransferase 1

Figure 2 Protein arginine methyltransferase 6 is methylated by protein arginine methyltransferase 1. A: Gel electrophoresis-autoradiographic image analysis of histone H3 methylation level change in the presence of protein arginine methyltransferase (PRMT) 1WT or PRMT1E153Q. 1 µM PRMT6 and 20 µM 14C-S-adenosyl methionine were incubated with or without 3 µM PRMT1WT, PRMT1E153Q or H3 protein respectively at 30 ℃ in the reaction buffer for 1h. The samples were separated on 12% sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis polyacrylamide gel. The radioactive SDS gel was dried and stored for two days and then scanned for the phosphor image; B: Western blot was performed to analysis PRMT1 level in HEK293T cells transfected by hPRMT1 or shScramble; C and D: Immunoprecipitation was performed to precipitate PRMT6 in HEK293T cell lysate transfected by shPRMT1 or shScramble or HA-PRMT1, followed by western blot to analysis monomethyl arginine and asymmetric dimethylarginine level of precipitated PRMT6. PRMT: Protein arginine methyltransferase.